The role and molecular mechanism of NOP16 in the pathogenesis of nasopharyngeal carcinoma.

We aimed to explore the effects of NOP16 on the pathogenesis of nasopharyngeal carcinoma (NPC) and the related mechanism. In this study, the expression level of NOP16 in NPC tissues and adjacent tissues was measured by qRT-polymerase chain reaction (PCR) and immunohistochemistry (IHC) tests. In the in vitro study, the expression levels of NOP16 and RhoA/phosphatidylinositol 3-kinase (PI3K)/Akt/c-Myc and IKK/IKB/NF-κB signalling pathway-related proteins in NPC cells were measured by qRT-PCR and Western blot (WB). CCK8 assays and colony formation assays were used to detect cell proliferation. Transwell assays were used to detect the migration and invasion ability of NPC cells. Flow cytometry and WB were used to measure the level of apoptosis. For the in vivo study, NPC xenograft models were established in nude mice, and tumour weight and volume were recorded. The expression levels of NOP16 and RhoA/PI3K/Akt/c-Myc signalling pathway-related proteins and mRNAs were measured by immunofluorescence, qRT-PCR and WB experiments. In clinical samples, the results of qRT-PCR and IHC experiments showed that the expression level of NOP16 was significantly increased in NPC tissues. In the in vitro study, the results of qRT-PCR and WB experiments showed that NOP16 was significantly increased in NPC cells. The CCK8 assay, colony formation assay, transwell assay and flow cytometry results showed that knocking out NOP16 inhibited the proliferation, migration and invasion of NPC cells and increased apoptosis. WB results showed that knocking out NOP16 inhibited the RhoA/PI3K/Akt/c-Myc and IKK/IKB/NF-κB signalling pathways. These effects were reversed by 740Y-P (PI3K activator). In the in vivo study, knockdown of NOP16 reduced tumour volume and weight and inhibited the RhoA/PI3K/Akt/c-Myc signalling pathway. In conclusion, knockdown of NOP16 inhibited the proliferation, migration and invasion of NPC cells and induced apoptosis by inhibiting the RhoA/PI3K/Akt/c-Myc and IKK/IKB/NF-κB pathways, leading to the malignant phenotype of NPC.

Whole-exome sequencing reveals the major genetic factors contributing to neuromyelitis optica spectrum disorder in Chinese patients with aquaporin 4-IgG seropositivity.

BACKGROUND AND OBJECTIVE: Neuromyelitis optica spectrum disorder (NMOSD) is an autoimmune disease. Although genetic factors are involved in its pathogenesis, limited evidence is available in this area. The aim of the present study was to identify the major genetic factors contributing to NMOSD in Chinese patients with aquaporin 4 (AQP4)-IgG seropositivity. METHODS: Whole-exome sequencing (WES) was performed on 228 Chinese NMOSD patients seropositive for AQP4-IgG and 1400 healthy controls in Guangzhou, South China. Human leukocyte antigen (HLA) sequencing was also utilized. Genotype model and haplotype, gene burden, and enrichment analyses were conducted. RESULTS: A significant region of the HLA composition is on chromosome 6, and great variation was observed in DQB1, DQA2 and DQA1. HLA sequencing confirmed that the most significant allele was HLA-DQB1*05:02 (p < 0.01, odds ratio [OR] 3.73). The genotype model analysis revealed that HLA-DQB1*05:02 was significantly associated with NMOSD in the additive effect model and dominant effect model (p < 0.05). The proportion of haplotype "HLA-DQB1*05:02-DRB1*15:01" was significantly greater in the NMOSD patients than the controls, at 8.42% and 1.23%, respectively (p < 0.001, OR 7.39). The gene burden analysis demonstrated that loss-of-function mutations in NOP16 were more common in the NMOSD patients (11.84%) than the controls (5.71%; p < 0.001, OR 2.22). The IgG1-G390R variant was significantly more common in NMOSD, and the rate of the T allele was 0.605 in patients and 0.345 in the controls (p < 0.01, OR 2.92). The enrichment analysis indicated that most of the genetic factors were mainly correlated with nervous and immune processes. CONCLUSIONS: Human leukocyte antigen is highly correlated with NMOSD. NOP16 and IgG1-G390R play important roles in disease susceptibility.

NOP16 promotes hepatocellular carcinoma progression and triggers EMT through the Keap1-Nrf2 signaling pathway.

BACKGROUND: Nucleolar protein 16 (NOP16) is present in the protein complex of the nucleolus. The NOP16 promoter contains a c-Myc binding site, and the transcriptional regulation by c-Myc directly regulates NOP16 expression levels. OBJECTIVE: Dysregulation of NOP 16 is currently reported in only a small number of cancers. In this study, the expression profile of NOP 16 in hepatocellular carcinoma (LIHC) and its clinical significance were analyzed. METHODS: NOP16 expression in hepatocellular carcinoma (LIHC) and its relationship with the clinical characters of LIHC were examined using the Cancer Genome Atlas (TCGA), the Gene Expression comprehensive database (GEO), Kaplan-Meier survival analysis, univariate Cox analysis, multivariate Cox analysis, ROC curve analysis of KEGG enrichment, GSEA enrichment, in vitro experiments (e.g., siRNA interference of NOP16 expression in hepatoma cells, Keap1-Nrf2 pathway, cell cycle, cell apoptosis and Transwell assays), and LIHC single-cell sequencing (scRNA). RESULTS: Pan-cancer analysis revealed that NOP16 was highly expressed in 20 cancer types, including LIHC, and high NOP16 expression was an independent adverse prognostic factor in LIHC patients. The expression levels of NOP16 mRNA and protein were significantly increased in tumour tissues of LIHC patients compared to normal tissues. The functions of co-expressed genes were primarily enriched in the cell cycle and reactive oxygen species metabolism. The experimental results showed that knockdown of NOP16 activated the Keap/Nrf2 signalling pathway and inhibited the invasion, migration, and EMT progression of LIHC cells. LIHC scRNA-seq data showed that NOP16 was primarily expressed in T lymphocytes. CONCLUSIONS: NOP16 promoted cancer development in LIHC and caused an imbalance in Keap/Nrf2 signalling, which subsequently caused the aberrant expression of genes typical for EMT, cell cycle progression and apoptosis. NOP16 is a potential prognostic marker and therapeutic target for hepatocellular carcinoma progression.

Elevated Expression of NOP16 as a Novel Prognostic Biomarker of Prostate Cancer.

OBJECTIVE: To investigate the role of NOP16, a target gene of estrogen and c-Myc, in prostate cancer (PCa). METHODS: The expressions of NOP16 in PCa and benign prostate hyperplasia (BPH) tissues, PC-3 and RWPE-1 cell lines were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot respectively. The expressions of NOP16 in BPH and PCa paraffin-embedded tissues were assessed by immunohistochemistry (IHC). The Cancer Genome Atlas (TCGA) database was used to analyze the effect of NOP16 gene expression on the disease-free survival of PCa patients. CCK-8, wound healing, transwell assays and flow cytometric analysis were used to assess the changes of proliferation, migration, invasion and apoptosis of PC-3 cells respectively after knocking down NOP16 in vitro. Gene ontology (GO) and gene set enrichment analysis (GSEA) were used to analyze the RNA sequencing data followed by knocking down NOP16 in PC-3 cells. Mouse models were used to explore the effect of NOP16 on PC-3 subcutaneous tumor growth in vivo. RESULTS: The NOP16 expression was significantly higher in PCa tissues than that in BPH tissues and significantly higher in PC-3 cells than in RWPE-1 cells. PCa patients with low NOP16 expression have a longer disease-free survival than that with high NOP16 expression. Ribosome biogenesis in eukaryotes, ECM-receptor interaction, PTEN and VEGF signaling pathways may be changed in the NOP16 knockdown group than control. Knockdown of NOP16 could significantly inhibit the proliferation, migration and invasion of PC-3 cells in vitro and in vivo. Low expression of NOP16 could reduce the total protein synthesis and induce the apoptosis of PC-3 cells. CONCLUSION: NOP16 may be expected to be a novel biomarker for predicting the occurrence and development of PCa, and may become a target for the treatment of PCa.

Caracterização da proteína nucleolar Nop 16 na fisiologia do patógeno Cryptococcus gattii

Atualmente, estima-se que cerca de 300 milhões de pessoas sejam acometidas por doenças fúngicas anualmente em todo o globo. Espécies de Aspergillus, Candida e Cryptococcus são os principais causadores de micoses que afetam, em especial, indivíduos imunossuprimidos. Espécies de C. neoformans e C. gattii se destacam como agentes causadores da criptococose, uma micose sistêmica e invasiva que pode atingir o sistema nervoso central e, se não tratada rapidamente, pode levar à morte. Em estudos anteriores, nosso grupo sugeriu que o gene NOP16 tem potencial para representar um alvo para agentes antifúngicos. Nesse estudo, buscamos entender o papel de Nop16 na fisiologia de C. gattii através da caracterização dos efeitos da deleção do gene NOP16 nas características fenotípicas e patogênicas de C. gattii. Análises por microscopia óptica, de fluorescência, eletrônica de varredura e de transmissão, combinadas com ensaios sorológicos e de interação com fagócitos, demonstraram que a deleção do gene acarreta sutis alterações no tamanho da cápsula polissacarídica, na taxa de fagocitose e na secreção de GXM. Entretanto, o estudo de vesículas extracelulares (EVs) por microscopia eletrônica de transmissão e por rastreamento de nanopartículas revelou que mutantes deficientes na expressão do gene NOP16 tiveram capacidade reduzida de produzir EVs. Esse fenótipo foi acompanhado de uma reduzida capacidade de levar à morte larvas de Galleria mellonella. Esses resultados indicam que Nop16 é um elemento importante na fisiologia e na patogênese de C. gattii.